LOCUS       sequence-338564-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion protein of Gluc and the light-activated 
            transcription factor EL222 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  AUTHORS   Adir O, Albalak MR, Abel R, Weiss LE, Chen G, Gruber A, Staufer O, 
            Kurman Y, Kaminer I, Shklover J, Shainsky-Roitman J, Platzman I, 
            Gepstein L, Shechtman Y, Horwitz BA, Schroeder A
  TITLE     Synthetic cells with self-activating optogenetic proteins 
            communicate with natural cells.
  JOURNAL   Nat Commun. 2022 Apr 28;13(1):2328. doi: 10.1038/s41467-022-29871-8.
  PUBMED    35484097
REFERENCE   2  (bases 1 to 6503)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-04-28"; volume: "13(1):2328. doi"; pages: " 
            10.1038/s41467-022-29871-8"
COMMENT     Sequence Label: pET28a-Gluc-EL222-his
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     955..974
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(994..1071)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1277..1296
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1380..1399
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1380..1398
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1399..1423
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1438..1460
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2680..2697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2747..2764
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2817..2835)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2831..2878
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2915..3370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3002..3021)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3212..3233
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3462..4277)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3462..4277)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4185..4204
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4269..4288
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4399..4987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4888..4907
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5141..5158
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5173..5312
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5238..5257)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5398..5420
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5414..5605)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6414..6503,1..993))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
       61 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
      121 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
      181 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
      241 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
      301 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
      361 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
      421 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
      481 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
      541 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
      601 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
      661 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
      721 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
      781 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
      841 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
      901 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
      961 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1021 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     1081 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     1141 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     1201 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     1261 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     1321 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     1381 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     1441 gtttaacttt aagaaggaga tatacatatg aaaccaacag aaaataacga agattttaac
     1501 atcgtcgcgg tagcctcaaa ttttgcaacc accgacttag acgcggatcg tgggaaatta
     1561 ccaggtaaga aattgccgtt ggaagtactt aaagaactgg aagccaacgc acgcaaggct
     1621 ggatgcactc gtgggtgttt gatctgtctg tcgcatatta agtgtacgcc aaagatgaaa
     1681 aagttcatcc cagggcgttg tcatacttac gaaggagaca aagagtcagc acagggcgga
     1741 atcggggaag ctatcgtaga tatccctgag atcccaggct tcaaagactt ggaaccgctg
     1801 gaacagttta ttgctcaagt tgacttgtgc gtcgactgta cgactgggtg ccttaaaggg
     1861 ctggcgaatg tccaatgtag tgatctgctt aagaagtggc ttcctcagcg ttgcgctaca
     1921 ttcgcaagca aaattcaggg gcaagtggat aagatcaaag gcgcgggtgg tgacggtggt
     1981 ggtggttcag gtggtggtgg ttcaatgttg gatatgggac aagatcggcc gatcgatgga
     2041 agtggggcac ccggggcaga cgacacacgc gttgaggtgc aaccgccggc gcagtgggtc
     2101 ctcgacctga tcgaggccag cccgatcgca tcggtcgtgt ccgatccgcg tctcgccgac
     2161 aatccgctga tcgccatcaa ccaggccttc accgacctga ccggctattc cgaagaagaa
     2221 tgcgtcggcc gcaattgccg attcctggca ggttccggca ccgagccgtg gctgaccgac
     2281 aagatccgcc aaggcgtgcg cgagcacaag ccggtgctgg tcgagatcct gaactacaag
     2341 aaggacggca cgccgttccg caatgccgtg ctcgttgcac cgatctacga tgacgacgac
     2401 gagcttctct atttcctcgg cagccaggtc gaagtcgacg acgaccagcc caacatgggc
     2461 atggcgcgcc gcgaacgcgc cgcggaaatg ctcaagacgc tgtcgccgcg ccagctcgag
     2521 gttacgacgc tggtggcatc gggcttgcgc aacaaggaag tggcggcccg gctcggcctg
     2581 tcggagaaaa ccgtcaagat gcaccgcggg ctggtgatgg aaaagctcaa cctgaagacc
     2641 agcgccgatc tggtgcgcat tgccgtcgaa gccggaatcc atcatcatca tcatcactaa
     2701 ggatccgaat tcgagctccg tcgacaagct tgcggccgca ctcgagcacc accaccacca
     2761 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2821 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2881 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2941 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3001 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3061 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3121 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3181 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3241 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3301 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3361 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3421 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     3481 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     3541 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     3601 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     3661 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     3721 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3781 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     3841 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     3901 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3961 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     4021 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     4081 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     4141 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     4201 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     4261 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     4321 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4381 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4441 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4501 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4561 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4621 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4681 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4741 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4801 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4861 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4921 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4981 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5041 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5101 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5161 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     5221 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     5281 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     5341 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5401 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     5461 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     5521 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     5581 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     5641 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     5701 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     5761 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     5821 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     5881 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     5941 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     6001 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     6061 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     6121 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     6181 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     6241 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     6301 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     6361 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     6421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6481 ggagaggcgg tttgcgtatt ggg
//
